Human prostatic cell line PNT1A, a useful tool for studying androgen receptor transcriptional activity and its differential subnuclear localization in the presence of androgens and antiandrogens
Human prostatic cell line PNT1A, a useful tool for studying androgen receptor transcriptional activity and its differential subnuclear localization in the presence of androgens and antiandrogens
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DOI：
10.1016/s0303-7207(01)00669-4
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发表时间：
2001-11-26
影响因子：
4.1
通讯作者：
Sultan, C
中科院分区：
文献类型：
--
作者：
Avancès, C;Georget, V;Sultan, C
人类永生化前列腺细胞系 PNT1A 已被证明是分析细胞过程（例如响应雄激素和生长因子的前列腺上皮增殖）的良好模型。在这里，我们通过分析抗雄激素的几种作用，使用该细胞系来研究雄激素受体（AR）的转录活性和运输。使用野生型人类 AR 和雄激素反应基因报告基因进行 PNT1A 细胞瞬时转染实验。我们证明，PNT1A 细胞以及前列腺癌细胞系 DU-145 中的天然雄激素（睾酮和二氢睾酮）可以触发报告基因的转录。在两种细胞系中进行竞争性实验。我们观察到10(-7) M的醋酸环丙孕酮(CPA)和羟氟他胺的拮抗能力没有差异。在此浓度下。比卡鲁胺拮抗剂活性较低。并联。我们比较了修饰的绿色荧光蛋白 (EGFP)-AR 在 COS-7 中的亚细胞定位。荧光显微镜下的 PNT1A 和 DU-145 细胞系：我们发现细胞核和细胞质之间的分布不同，具体取决于细胞系和培养基。雄激素诱导 PNT1A 和 DU-145 细胞核内簇的形成。然而，在同一活细胞和核灶中雄激素结合 EGFP-AR 的细胞核运输在 PNT1A 细胞中更容易检查。比卡鲁胺的抗雄激素能力通过 EGFP-AR 较慢的雄激素依赖性核转移和均匀的核定位来体现。在 CPA 存在的情况下观察到核簇出现延迟，我们得出结论，PNT1A 细胞系是比 DU-145 细胞系更好的模型，可以分析 AR 的运输和 AR 在核基质上的关联，以及观察抗雄激素对前列腺细胞中这些关键步骤的作用。 (C) 2001 年，爱思唯尔科学爱尔兰有限公司出版。
The human immortalized prostatic cell line PNT1A has been proved to be a good model for analysis of cellular processes such as the prostatic epithelium proliferation in response to androgens and growth factors. Here we used this cell line for studying the transcriptional activity and trafficking of the androgen receptor (AR) by analyzing several actions of antiandrogens. Transient transfection experiments with PNT1A cells were performed with wild type human AR and an androgen-responsive gene reporter. We demonstrated that the transcription of reporter gene could be triggered by natural androgens (testosterone and dihydrotestosterone) in PNT1A cells as well as in the prostatic carcinoma cell line DU-145. With competitive experiments in the two cell lines. we observed no difference between the antagonistic capacity of cyproterone acetate (CPA) and hydroxyflutamide at 10(-7) M. At this concentration. bicalutamide antagonist activity was lower. In parallel. we compared the subcellular localization of the modified green fluorescent protein (EGFP)-AR in COS-7. PNT1A and DU-145 cell lines under fluorescence microscopy: we found different distributions between nucleus and cytoplasm, depending on the cell line and the culture medium. Androgen induced cluster formation within the nucleus of the PNT1A and DU-145 cells. However, the cytonuclear trafficking of androgen bound EGFP-AR in the same living cell and nuclear foci were easier to examine in the PNT1A cells. The antiandrogen capacity of bicalutamide was manifested by a slower androgen-dependent nuclear transfer of EGFP-AR and a homogeneous nuclear localization. A delayed advent of nuclear clusters was observed in presence of CPA, We conclude that the PNT1A cell line is a better model than the DU-145 cell line to analyze the trafficking of AR and the association of AR on the nuclear matrix, as well as to observe the action of antiandrogens on these critical steps in prostate cells. (C) 2001 Published by Elsevier Science Ireland Ltd.